EP0217768B1 - Méthode et composés pour la mise en évidence de l'héparine - Google Patents
Méthode et composés pour la mise en évidence de l'héparine Download PDFInfo
- Publication number
- EP0217768B1 EP0217768B1 EP86850288A EP86850288A EP0217768B1 EP 0217768 B1 EP0217768 B1 EP 0217768B1 EP 86850288 A EP86850288 A EP 86850288A EP 86850288 A EP86850288 A EP 86850288A EP 0217768 B1 EP0217768 B1 EP 0217768B1
- Authority
- EP
- European Patent Office
- Prior art keywords
- heparin
- plasma
- factor
- admixture
- blood
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/56—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/86—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood coagulating time or factors, or their receptors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
- G01N2333/964—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue
- G01N2333/96425—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals
- G01N2333/96427—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general
- G01N2333/9643—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general with EC number
- G01N2333/96433—Serine endopeptidases (3.4.21)
- G01N2333/96441—Serine endopeptidases (3.4.21) with definite EC number
- G01N2333/96444—Factor X (3.4.21.6)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2400/00—Assays, e.g. immunoassays or enzyme assays, involving carbohydrates
- G01N2400/10—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- G01N2400/38—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence, e.g. gluco- or galactomannans, e.g. Konjac gum, Locust bean gum, Guar gum
- G01N2400/40—Glycosaminoglycans, i.e. GAG or mucopolysaccharides, e.g. chondroitin sulfate, dermatan sulfate, hyaluronic acid, heparin, heparan sulfate, and related sulfated polysaccharides
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2405/00—Assays, e.g. immunoassays or enzyme assays, involving lipids
- G01N2405/04—Phospholipids, i.e. phosphoglycerides
Definitions
- This invention pertains to an improved method for determining the amount of heparin in a blood sample, and compositions useful in connection with the method.
- Heparin establishes an important parameter for the supervision of heparin treatment which is often administered in the presence of a threat of thrombosis.
- Heparin forms with antithrombin III (AT III) a complex which inhibits the proteolytic activity of thrombin. Since thrombin catalyzes the formation of fibrin from fibrinogen,the thrombin activity is responsible for the coagulation of blood or plasma and hence also for the fibrin clots which form in thrombosis.
- Heparin treatment is often applied in the presence of a threat of thrombosis (e.g. before surgical interventions). A precise adjustment of heparin concentration is therefore extremely important.
- the patient's plasma sample is incubated with normal plasma, buffer, and a known excess of Factor Xa for a predetermined time period, after which a sub sample from this primary reaction mixture is removed and assayed for Factor Xa activity.
- the latter step constitutes the second-stage of the assay.
- Factor Xa activity is measured by the addition of the test sample to another test tube, and to it calcium chloride, cephalin in bovine plasma are added separately in timed fashion. commercial embodiment of this method is described in detail in Sigma Chemical Company Technical Bulletin No. 870 (as revised in January 1982).
- step (b) comparing the clotting time in step (b) with the clot times on an established calibration curve obtained by plotting clotting time versus heparin concentration to thereby determine the amount of heparin in the patient blood plasma sample.
- the present invention involves a method for determining the concentration of heparin in a blood plasma sample, which method comprises
- the method is suitably carried out with 100 microliters of a patient's blood plasma, 100 microliters of Factor X a and 100 microliters of the admixture set forth in (B) above.
- the Factor X a can be prepared from bovine plasma by any method described in the literature. For example, Yin, E.T., Wessler, S., and Stoll, P., J. Biol. Chem. 243: 112 (1968). The purified Factor X a is further lyophilized and stabilized in a buffer containing crystalline bovine serum albumin, polyethylene glycol, NaCl, and Tris (hydroxymethyl) aminomethane maleate, pH 7.5. Factor X a as described in the aforementioned Sigma-Bulletin No. 870 can be used.
- Incubation of the patient's blood plasma and Factor X a preferably takes place at 37°C for 60-160 seconds, and more preferably for a time period of 120 seconds.
- the plasma fraction of (B) (3) is mammalian blood that is substantially free of Factors II, VII, IX and X but which still contains Factor V and fibrinogen, and can be produced by the steps of
- the buffered plasma fraction produced by steps (i)-(v) is admixed with calcium chloride and brain phospholipids and the resulting admixture has the characteristics (a)-(d) set forth above.
- step (i) 9 volumes of the mammalian blood can be treated with one volume of 3.8% sodium citrate solution to effect anti-coagulation and then centrifuged at 2500 x g to remove blood cells. Bovine blood is preferred.
- step (ii) the separation can be effected by adding to 1000 ml of plasma from step (i) 7.35 g of trisodium citrate, stirring to dissolve the citrate, followed by the addition of 20.825 g of barium chloride (sufficient to achieve an overall barium chloride concentration of about 0.1 M in the plasma mixture) at room temperature over a period of 1-2 hours, and the heavy precipitate thus formed with the aid of barium citrate can be removed by the centrifugation of the plasma mixture at 3000 x g for 20 minutes (the heavy precipitate containing Factors II, VII, IX and X) leaving a clear plasma supernatant.
- the separation can take the form of fractionation with solid ammonium sulfate at a concentration of approximately 270 g per liter of the plasma at room temperature. If the resulting plasma mixture is allowed to sit at room temperature for 1-2 hours, and then centrifuged at 4000 x g for 15 minutes at room temperature, a precipitated protein fraction can be obtained.
- concentration of ammonium sulfate of 270 g/l corresponds to a 40-45% saturated solution.
- a salt concentration of about 30% saturation fibrinogen precipitates out; as the salt concentration increases above 30% saturation to the area of 40% saturation the Factor V proteins precipitate out, as well as some other as yet unidentified proteins.
- ammonium sulfate concentration of 270 g/l is quite suitable in precipitating out fibrinogen and Factor V proteins
- ammonium sulfate to precipitate fibrinogen is described in a number of publications, e.g. "The Biochemistry of Blood Coagulation” by T. Astrup, in Acta Physiologica Scandanavia, Supplement 21,1944.
- ammonium sulfate for the fractionation of Factor V is also described in a number of publications, e.g. the publication "Human Blood Coagulation and Its Disorders" edited by Biggs & MacFarlane, p. 54, 3rd edition 1962.
- step (iv) the protein precipitate from step (iii) is harvested and dissolved in a volume of distilled water equal to 30-40% of the original plasma volume.
- This "plasma fraction” is then dialysed against 0.9% NaCl solution until no ammonium ion is present in the fraction.
- the dialysed plasma fraction is clarified by centrifugation at 3000 x g for 15 minutes at room temperature.
- step (v) the-clear plasma fraction from step (iv) can be buffered to a pH of 7.5 by mixing nine volumes of it with one volume of a solution containing 2.50 g polyethylene-glycol, 90.0 g NaCl and 98.88 g Tris-maleate per liter of distilled water.
- the equipment that is recommended includes 12 ⁇ 75 mm glass test tubes, pipettes to deliver 0.1 ml, distilled water, two stopwatches and a 37°C water bath. If a clot detector machine is to be used, such as a BBL Fibrometer, a 0.3 ml probe is employed.
- step 1-3 The number of seconds it takes for the above mixture (steps 1-3) to form a solid clot is noted and can be converted to units of heparin per ml of plasma using a standard heparin calibration curve such as is shown in Figure 1.
- the methods for preparing such calibration curves are well known in the art (e.g. the aforementioned Sigma Technical Bulletin 870).
- the dialysed plasma fraction was then clarified by centrifugation at 3000 x g for 15 minutes at room temperature and then buffered to a pH of 7.5 by mixing nine volumes of it with one volume of a solution containing 2.5 g of polyethylene glycol, 90.0 g of NaCl and 98.88 g of tris-maleate, dissolved in 1 liter of distilled water.
- Calcium chloride was added to this buffered plasma fraction to achieve a calcium chloride concentration of 0.025 M, whereafter cephalin [prepared by Bell and Alton, Nature , 174: 880 (1954)] was added in a volume ratio of one volume of such cephalin to 99 volumes of the buffered plasma fraction to which calcium chloride had already been added.
- cephalin prepared by Bell and Alton, Nature , 174: 880 (1954)
- the resulting admixture served as one of the assay materials for the method and Factor X a served as the other assay material for the method.
- Example 2 The procedure of Example 2 was followed except that the Factor X a used was Activated Factor X Reagent, Stock No. 870-10 from the Sigma Chemical Company of St. Louis, Missouri. Results comparable to that of Example 2 were obtained.
- Example 2 In order to decrease the clotting time in Example 2 the concentration of cephalin in Example 1 can be increased or the Factor X a concentration in Example 2 can be increased.
- the invention can be made available in kit form that would comprise in a freeze-dried form, (A) a container of Factor X a and (B) a container of an admixture of calcium chlroide, brain phospholipids and a buffered plasma fraction that has been produced by treating mammalian blood to substantially remove clotting factors II, VII, IX and X.
- the components (A) and (B) are preadjusted so that when employed in the assay they will provide optimal sensitivity to the heparin in blood plasma samples.
- heparin includes heparin and heparin-like compounds whether they be natural, synthetic, high or low molecular weight heparin fractions or fragments.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Urology & Nephrology (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Cell Biology (AREA)
- Neurosurgery (AREA)
- Biophysics (AREA)
- General Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Pathology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Claims (5)
- ledit mélange étant caractérisé par le fait que
- ledit mélange étant caractérisé par le fait que
- ledit mélange étant caractérisé par le fait que
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AT86850288T ATE64470T1 (de) | 1985-09-05 | 1986-09-02 | Verfahren und zusammensetzungen zur bestimmung von heparin. |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US77284685A | 1985-09-05 | 1985-09-05 | |
US772846 | 1985-09-05 |
Publications (3)
Publication Number | Publication Date |
---|---|
EP0217768A2 EP0217768A2 (fr) | 1987-04-08 |
EP0217768A3 EP0217768A3 (en) | 1988-12-21 |
EP0217768B1 true EP0217768B1 (fr) | 1991-06-12 |
Family
ID=25096437
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP86850288A Expired - Lifetime EP0217768B1 (fr) | 1985-09-05 | 1986-09-02 | Méthode et composés pour la mise en évidence de l'héparine |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP0217768B1 (fr) |
AT (1) | ATE64470T1 (fr) |
DE (1) | DE3679763D1 (fr) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102005028018A1 (de) * | 2005-06-16 | 2006-12-21 | Dade Behring Marburg Gmbh | Verfahren zur Standardisierung von Gerinnungstesten |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1989008150A1 (fr) * | 1988-03-03 | 1989-09-08 | Nippon Shoji Kabushiki Kaisha | Procede de mesure de l'activite biologique de l'antithrombine iii et reactifs de mesure |
FR2640385B1 (fr) * | 1988-12-08 | 1991-02-08 | Serbio | Nouveau procede de dosage de l'heparine et son utilisation dans les trousses de dosage |
US5308755A (en) * | 1992-06-08 | 1994-05-03 | Research Corporation Technologies, Inc. | Method for measuring heparin |
DK0825443T3 (da) * | 1996-08-17 | 2001-08-06 | Aventis Behring Gmbh | Fremgangsmåde til mængdebestemmelse af glycosaminoglycaner i antitrombin III-holdige opløsninger |
GR1002776B (el) * | 1996-12-02 | 1997-09-26 | . | Εξουδετερωση της αντιπηκτικης δρασης του συμπλεγματος της αντιθρομβινης-ιιι/ ηπαρινης, νεα δοκιμασια αποκαλυψης θρομβοφιλικων καταστασεων |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
NL7602423A (nl) * | 1976-03-08 | 1977-09-12 | Leuven Res & Dev Vzw | Bepaling van heparine in bloedplasma. |
US4067777A (en) * | 1976-05-13 | 1978-01-10 | Innerfield Irving | Determination of heparin in the blood |
DE2812943C3 (de) * | 1978-03-23 | 1981-05-14 | Boehringer Mannheim Gmbh, 6800 Mannheim | Verfahren und Reagens zur Bestimmung der biologischen Aktivität von Heparin im Plasma |
DE3005540A1 (de) * | 1980-02-14 | 1981-08-20 | Boehringer Mannheim Gmbh, 6800 Mannheim | Verfahren und reagens zur bestimmung der biologischen aktivitaet von heparin im plasma |
-
1986
- 1986-09-02 AT AT86850288T patent/ATE64470T1/de not_active IP Right Cessation
- 1986-09-02 EP EP86850288A patent/EP0217768B1/fr not_active Expired - Lifetime
- 1986-09-02 DE DE8686850288T patent/DE3679763D1/de not_active Expired - Lifetime
Non-Patent Citations (1)
Title |
---|
CHEMICAL ABSTRACTS, vol. 93, no. 3, 21 July 1980, Columbus, OH (US); E.T.YIN et al., p. 337, no. 22057w * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102005028018A1 (de) * | 2005-06-16 | 2006-12-21 | Dade Behring Marburg Gmbh | Verfahren zur Standardisierung von Gerinnungstesten |
Also Published As
Publication number | Publication date |
---|---|
EP0217768A3 (en) | 1988-12-21 |
ATE64470T1 (de) | 1991-06-15 |
DE3679763D1 (de) | 1991-07-18 |
EP0217768A2 (fr) | 1987-04-08 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Yin et al. | Plasma heparin: a unique, practical, submicrogram-sensitive assay | |
US4946775A (en) | Composition, kit and method for assaying heparin and a method for making the composition | |
EP0482088B1 (fr) | Regulations ameliorees pour effectuer une coagulation stable du sang | |
EP1092157B1 (fr) | Regulateurs de la coagulation pour des essais pt et aptt | |
EP0706658B1 (fr) | Plasma d'etalonnage pour tests de temps de prothrombine | |
US4948724A (en) | Composition, kit and method for assaying heparin and a method for making the composition | |
Osterud | How to measure factor VII and factor VII activation | |
EP0360871B2 (fr) | Procede de mesure de l'activite biologique de l'antithrombine iii et trousse d'essai pour sa mesure | |
US7294479B2 (en) | Compositions, kit and one-step method for monitoring compounds having anti-factor Xa and/or anti factor IIa activities | |
EP0217768B1 (fr) | Méthode et composés pour la mise en évidence de l'héparine | |
US4851336A (en) | Composition, kit, and method for assaying heparinano a method for making the composition | |
Corrigan Jr et al. | Factor II antigen in liver disease and warfarin‐induced vitamin K deficiency: Correlation with coagulant activity using echis venom | |
US5308756A (en) | Protein S chromogenic assay | |
US5221614A (en) | Method and reagent for determining the biological activity of antithrombin III by measuring coagulation time | |
EP0406971B1 (fr) | Méthode et trousse de réactifs pour déterminer l'activité fonctionnelle de la protéine S dans le plasma humain | |
Nishida et al. | A new rapid and simple assay for factor XIII activity using dansylcadaverine incorporation and gel filtration | |
JPH0235942B2 (fr) | ||
Duncan et al. | A clinical evaluation of automated chromogenic tests as substitutes for conventional prothrombin time and activated partial thromboplastin time tests. | |
US5476771A (en) | Test for quantitative thrombin time | |
Nyman | The preparation of an artificial reagent for the one-stage factor VIII assay | |
CA2137342A1 (fr) | Test quantitatif pour la determination du temps de thrombine | |
Kandall et al. | Determinants of Prothrombinase Activity and Modification of Prothrombin Conversion by Thrombin‐Treated Factor V | |
EP0186476A2 (fr) | Trousse de diagnostic et procédé pour le dosage de composants sanguins | |
Kirchhof et al. | Control of anticoagulant therapy with a chromogenic substrate | |
Grannis et al. | The spectrophotometry determination of thrombin activity in plasma |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
AK | Designated contracting states |
Kind code of ref document: A2 Designated state(s): AT DE FR GB IT NL SE |
|
PUAL | Search report despatched |
Free format text: ORIGINAL CODE: 0009013 |
|
AK | Designated contracting states |
Kind code of ref document: A3 Designated state(s): AT DE FR GB IT NL SE |
|
17P | Request for examination filed |
Effective date: 19890615 |
|
17Q | First examination report despatched |
Effective date: 19900528 |
|
GRAA | (expected) grant |
Free format text: ORIGINAL CODE: 0009210 |
|
AK | Designated contracting states |
Kind code of ref document: B1 Designated state(s): AT DE FR GB IT NL SE |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: NL Effective date: 19910612 Ref country code: AT Effective date: 19910612 |
|
REF | Corresponds to: |
Ref document number: 64470 Country of ref document: AT Date of ref document: 19910615 Kind code of ref document: T |
|
REF | Corresponds to: |
Ref document number: 3679763 Country of ref document: DE Date of ref document: 19910718 |
|
ITF | It: translation for a ep patent filed |
Owner name: DR. ING. A. RACHELI & C. |
|
ET | Fr: translation filed | ||
NLV1 | Nl: lapsed or annulled due to failure to fulfill the requirements of art. 29p and 29m of the patents act | ||
PLBE | No opposition filed within time limit |
Free format text: ORIGINAL CODE: 0009261 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT |
|
26N | No opposition filed | ||
EAL | Se: european patent in force in sweden |
Ref document number: 86850288.1 |
|
REG | Reference to a national code |
Ref country code: GB Ref legal event code: IF02 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: GB Payment date: 20040922 Year of fee payment: 19 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: DE Payment date: 20040923 Year of fee payment: 19 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: FR Payment date: 20040927 Year of fee payment: 19 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: SE Payment date: 20040928 Year of fee payment: 19 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: IT Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES;WARNING: LAPSES OF ITALIAN PATENTS WITH EFFECTIVE DATE BEFORE 2007 MAY HAVE OCCURRED AT ANY TIME BEFORE 2007. THE CORRECT EFFECTIVE DATE MAY BE DIFFERENT FROM THE ONE RECORDED. Effective date: 20050902 Ref country code: GB Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20050902 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: SE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20050903 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: DE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20060401 |
|
EUG | Se: european patent has lapsed | ||
GBPC | Gb: european patent ceased through non-payment of renewal fee |
Effective date: 20050902 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: FR Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20060531 |
|
REG | Reference to a national code |
Ref country code: FR Ref legal event code: ST Effective date: 20060531 |